Smith D D, Pratt K A, Sumner I G, Henneke C M
Department of Protein Engineering, Reading Laboratory, UK.
Protein Eng. 1995 Jan;8(1):13-20. doi: 10.1093/protein/8.1.13.
A protein designed de novo to fold into the Greek key jellyroll structural motif has been studied. Theoretical analyses have indicated that the designed sequence should adopt the beta-strand arrangement of the Greek key jellyroll rather than any other arrangement. A synthetic gene was constructed and the protein expressed in Escherichia coli. Circular dichroism spectroscopy is consistent with the protein folding into the designed conformation and also suggests the presence of tertiary structure. Fluorescence spectroscopy showed the single tryptophan to be partially buried, while denaturation studies showed changes in fluorescence to precede alterations in secondary structure.
对一种全新设计的、可折叠成希腊钥匙果冻卷结构基序的蛋白质进行了研究。理论分析表明，所设计的序列应采用希腊钥匙果冻卷的β-链排列方式，而非其他排列方式。构建了一个合成基因，并在大肠杆菌中表达了该蛋白质。圆二色光谱表明该蛋白质折叠成了设计的构象，同时也暗示了三级结构的存在。荧光光谱显示单一色氨酸被部分掩埋，而变性研究表明荧光变化先于二级结构的改变。
